Miscibility and Thermal Study of 4-Hydroxycoumarin Doped ...

RESEARCH ARTICLE Am. J. PharmTech Res. 2021; 11(1) ISSN: 2249-3387

Please cite this article as: Chougale RB et al., Miscibility and Thermal Study of 4-Hydroxycoumarin

Doped Chitosan Films. American Journal of PharmTech Research 2021.

Miscibility and Thermal Study of 4-Hydroxycoumarin Doped

Chitosan Films

Vinayak N. Vanjeri1, Naganagouda Goudar1, Saraswati P. Masti2, Ravindra B. Chougale1*

1.P. G. Department of Studies in Chemistry, Karnatak University, Dharwad-580 003, Karnataka,

India.

2.Department of Chemistry, Karnatak Science College, Dharwad - 580 001, India

ABSTRACT

In this study, new Chitosan/4-Hydropxycoumarin films were prepared and characterized. The

influence of 4-Hydroxycoumarin on the surface morphology, thermal behavior of the chitosan

films were studied. The experimental studies showed that surface morphology becomes uniform

and surface roughness increases with increasing the concentration of 4-Hydroxycoumarin in the

chitosan film. The appreciable intermolecular interaction among the chitosan and 4-

Hydroxycoumarin is confirmed by the FTIR study. The thermal properties of the chitosan films

slightly increased with the increasing concentrations of 4-Hydroxycoumarin. Also, the presence of

single glass transition temperature in all Chitosan/4-Hydroxycoumarin films suggests the

components present in the film were miscible. Due to the existence of 4HC into the chitosan film,

the crystalline nature and water contact angle of CS decreases for the C4HC films. It can be

expected that, the best properties of Chitosan/4-Hydroxycoumarin films were recorded in the study

may play a vital role in food packaging and coating applications.

Keywords: Surface morphology, thermal behaviour, glass transition temperature, miscibility

*Corresponding Author Email: [email protected] Received 02 January 2021, Accepted 29 January 2021

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Chougale et. al., Am. J. PharmTech Res. 2021;11(1) ISSN: 2249-3387

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INTRODUCTION

In recent years, biodegradable polymers including cellulose, starch, chitosan and natural gums

have been studied extensively to produce potential biodegradable packaging material with

enhanced physicochemical properties. Nowadays there is an urgency to manufacture biodegradable

and environmental friendly bio-based polymeric material 1 for active food packaging. Active food

packaging is made to interact with product or its environment to extend shelf life of food. The

basic raw materials for film forming and coatings can be obtained from the natural sources

including starch, cellulose, proteins, polysaccharides, lipids and resins 2 which act as excellent

barrier to oxygen, water vapour and oil 3.

Among natural biopolymers, chitosan (CS) is a natural cationic polysaccharide with active amino

functional groups. CS is the deacetylated product of chitin which found most abundant

polysaccharide in nature 4-5. CS is a copolymer of N-acetyl-D-glucosamine and D-glucosamine.

Besides its biodegradability and biocompatibility, CS has been widely reported that it has proven

to have good antimicrobial property 6 against bacteria, yeasts and fungi 7-8. Several approaches

have been undertaken to get over the limitations of CS including polymerization and blending with

other polymers which paid much attention to alter or tailoring the property of interest. CS, its own

or blend component is used as a biomaterial 9-11 in water treatment, in food packaging and

medicine 12-19.

4-Hydroxycoumarins (4HC) is one of the important precursors in the realm of organic synthesis.

The interest towards 4HC has been amplified because, it is not only significant synthetic endpoints

20-21, but it contains structural nucleus of many natural products 22-24. The derivatives of 4HC have

shown a remarkably applications in pharmacological and physiological activities. The derivatives

of 4HC are used as anticoagulant, antibacterial, antifungal, antitumor, antioxidant, anti-

inflammatory agents 25-31. Also, in recent years there are references to derivatives with HIV

protease inhibitors 32. In addition, 4HC is an important fungal metabolite and its production leads

to further fermentative production of the natural anticoagulant dicoumarol. The dicoumarol is a

fermentation product found in spoiled sweet clover silages and is considered a mycotoxin 33. The

study aims to prepare 4HC doped CS films. Also the present work intended to explore the

influence of 4HC on CS films.

MATERIALS AND METHOD

Materials

The materials used in this study are chitosan from shrimp shells 75% (deacetylated) with viscosity

min 200 cps was purchased from Loba Chemie Pvt. Ltd, Mumbai, India. 4-Hydroxycoumarin


Chougale et. al., Am. J. PharmTech Res. 2021; 11(1) ISSN: 2249-3387

47 www.ajptr.com

(Sigma Aldrich), Acetic acid (Spectrochem Pvt. Ltd. Mumbai. India) and millipore water was

used.

Preparation of the Chitosan Films

CS films were prepared by doping different concentration of 4HC onto the CS solution using

solvent casting method and the composition for CS and CS/4HC films is shown Table 1. An

exactly weighed (2 g) amount of CS was dissolved in 150 mL of 2% acetic acid. To the CS

solution different concentrations of 4HC (0.02 g to 0.08 g dissolved in 5 mL 100% acetic acid)

were mixed and the mixture of CS and 4HC (C4HC) was stirred for 3-4 h. Then, subsequently,

definite volumes of homogeneous C4HC film solutions were poured onto the previously cleaned

and dried petri dishes and left for solvent evaporation at normal room temperature for a couple of

weeks. After ensuring evaporation of solvent, the films were peeled from petri dishes and stored in

vacuum desiccators for further characterization.

Table 1: Composition table of CS and CS/4HC films.

Sample Code Wt of Chitosan Wt of 4HC

CS 2 g 00 g

C4HC-1 2 g 0.02 g

C4HC-2 2 g 0.04 g

C4HC-3 2 g 0.06 g

C4HC-4 2 g 0.08 g

CHARACTERIZATIONS

Atomic Force Microscopy (AFM)

The surface morphology of the composite film was recorded using Atomic force microscopy

(AFM) using Nanosurf Easyscan2, (Switzerland) with the aluminum coated cantilever. All the

topographic images of film samples were collected in contact angle mode using aluminum coated

cantilever. The topographic images of film samples were taken and roughness of the films was

analyzed.

Fourier Transform Infrared (FTIR) Spectroscopy

The Fourier transform infrared spectroscopy was used to probe the interaction among the

components and films samples were screened for interaction using an ATR (attenuated total

reflection) method of IR spectrometer (Perkin-Elmer Spectrum Version 10.5.4). All the film

specimens were scanned between the 550 cm-1 to 4000 cm-1 at 4 cm-1 resolution.

Differential Scanning Calorimetry (DSC)

The DSC measurements were carried by using DSC Q20-V24.4 Build 122 system (TA

Instruments, USA). The instrument has balance sensitivity and in the alumina pans samples were



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loaded and reference pan kept empty for film sample analyis. In addition, pan was heated under

nitrogen atmosphere and a heating rate of the sample was kept 10oC/min.

Thermogravimetric Analysis (TGA)

A thermogravimetric analysis technique (SDT Q600 V20.9 Build 20 –Universal V4.5A TA

Instruments) was used to provide weight loss which is useful for the study of thermal stability of

films. The film samples of masses 5 to 6 mg were used and heated in an inert nitrogen atmosphere

(heating rate of 10°C/min) from ambient temperature to 600◦C. The weight losses at different

stages were analyzed from the curves of TGA.

The X-ray Diffraction (XRD)

XRD analysis of the films was carried out using a Rigaku SmartLab (Tokyo, Japan) X-ray

diffractometer. A Cu K-beta radiation was used with working voltage 40 kV and current 30 mA.

The scan was performed with continuous mode in the 2θ range from 5ᴼ to 80ᴼ and speed was 5ᴼ

min−1.

Water Contact Angle (WCA) Measurements

The water contact angles of the films were measured by the drop method using a contact angle

meter Model DMs-401 (Kyowa Interface Science Co. Ltd., Tokyo) to examine the hydrophilicity.

A drop of millipore water was carefully dropped on the film surface, and the contact angles were

measured. Each reported contact angle is the average value of three measurements.

RESULTS AND DISCUSSION

Atomic Force Microscopy

The surface morphology of the pure CS and C4HC films were analyzed by using atomic force

microscopy and obtained topographic images with their 3D view were shown in Figure 1. For the

topographic image of CS, 3.37 mV roughness was found. The results of AFM study indicates that,

the roughness of C4HC films (0.02-0.08 g 4HC) was increase when compared to that of pure CS

film, begins to alter roughness. This could be due to the distribution of 4HC in CS which

influenced on viscosity of the CS film solution. After addition of 4HC, the area roughness slightly

increased which confirmed that 4HC is less compatible with CS.



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Figure 1: AFM topographic images and their 3D views.

Fourier Transform Infrared Spectroscopy

Fourier transform infrared spectroscopic analysis was carried to confirm the interaction among

components. The FTIR spectra of CS and C4HC films are shown in Figure 2. The spectra of pure

chitosan film shows a broad band at 3365 cm-1 which is due to the OH and NH hydrogen band

stretching, & 2852 cm-1 CH stretching, 1642 cm-1 amide-I and 1029 cm-1 CO stretching vibration.

The band at 1558 cm-1 is assigned for the NH bending (amide-II). The bands at 2921, 1412 and



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1316 cm-1 are assigned to CH2 bending due to pyranose ring. The FTIR spectra of 4HC shows

peak at 3355 cm-1 which is due to the OH stretching and 1705–1599 cm-1 attributed to the –C=O

vibrational stretching. The peak observed at 2981 attributed to the aromtatic –C-H stretching. The

FTIR spectra of C4HC composite films showed changes in the peak value indicating that

interaction among the CS and 4HC. The –OH observed in the CS is shifted to the lower value

(3365 cm-1 to 3257 cm-1) in C4HC films. This might be due to the hydrogen bonding in the films

which leads to the intermolecular interaction. Also peak observed in the CS (2921 cm-1) appeared

at higher level and peak appeared at 1642 cm-1 shifted to the lower level (1636 cm-1) in C4HC

films. The shift in the peak value and peak intensity confirms that there is considerable interaction

among the components.

Figure 2: FTIR spectra of CS and C4HC films.

Thermogravimetric Analysis

The stability of the films were evaluated by using thermogravimetric analysis (TGA). Thermal

distraction of chitosan presented two significant weight losses as shown in Figure 3. Initial weight

loss observed at 41oC to 75oC due to loss of moisture and bound water (19.52 %). The second

major weight loss of 49.57 % was observed at 259oC to 325oC, this could be attributed to the

decomposition of saccharide structure present in the chitosan. Also, the incorporation of 4HC onto

the CS presented two step degradation patterns. The increased thermal stability observed in the

C4HC films. It is worth noting that, remarkable changes were noticed in C4HC series. The initial

weight loss observed from 39oC to 156oC, this could be due to the evaporation of physically

bonded water molecules. The maximum weight loss with 50 % to 54 % was observed between the



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temperature ranges 258oC to 330oC which is considerably higher than pure CS. This could be due

to the fully destruction of films. The strong intermolecular interaction leads to the increased

thermal stability of the film. This fact could indicate a good interaction between CS and 4HC. The

results of TGA were good agreement with FTIR study.

Figure 3: Thermogram of CS and C4HC films.

Differential Scanning Calorimetry

The graphs of DSC thermograms for CS and C4HC films were shown in the Figure 4. The

importance of Tg can be realized in the study of miscibility of components. Moreover, the

miscibility of the films depends upon the composition and the solvent used. In the C4HC films, Tg

significantly decreased to lower value 45.39oC, 48.79oC, 49.99 oC and 53.39 oC for C4HC-1,

C4HC-2, C4HC-3 and C4HC-4 respectively, this could be due to the considerable interaction

among the CS and 4HC which indicates composite films were miscible.

Figure 4: DSC Thermogram of CS and C4HC films.



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X-ray Diffraction study

The X-ray diffraction patterns of CS and C4HC films are shown in Figure 5. Two crystalline peaks

at 2θ = 11.59º and 28.9º were observed in CS film, these observations agreed with the results

reported by others 34-35. After the addition of 4HC in the chitosan film, the intensity of diffraction

peak 11.59º of CS is diminished with increasing content of 4HC. It illustrates that the crystalline

nature of CS decreases for the C4HC films due to the existence of 4HC. Also, the absence of any

new diffraction peaks for C4HC films reveals a complete dissociation of 4HC on the CS matrix.

Figure 5: X-ray diffraction patterns of CS and C4HC films.

Water Contact Angle Measurement

Water contact angle measurements were carried out to understand the hydrophilic nature of the

films. Below Figure 6 shows the images of water drops on the surface of films with contact angles.

It is well known that, when contact angle values greater than 90° are obtained, there are

hydrophilic interactions between the solid surface and the dissolution medium. The water contact

angles 83.9ᴼ, 88.7ᴼ, 86.3ᴼ, 79.7ᴼ and 74.3ᴼ were found for CS, C4HC-1, C4HC-2, C4HC-3, and

C4HC-4 respectively. It implies that all the films were hydrophilic. After addition of 4HC into the

CS the decreased hydrophilicity was observed for C4HC-1 film, further increase in the

concentration of 4HC the contact angles were decreased gradually. These results were correlated

with the AFM results that the contact angles were decreased with increasing the roughness of the

films. This behavior is likely to be associated with the hydrophobic backbone of the polymer

chains. This effect may be due to the fact that the interaction between 4HC and CS.



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Figure 6: Water drop images on CS and C4HC films.

CONCLUSION

In the present study CS and C4HC films were successfully prepared and characterized. The results

of AFM study showed the uniform surface morphology and surface roughness showed increasing

order as the weight of the 4HC is increased in the CS film. The compatibility among components

indicates the appreciable intermolecular interaction among the CS and 4HC which is confirmed by

the FTIR study. The thermal properties of the CS films slightly increased with incorporation of

different concentration of 4HC. In addition presence of single glass transition temperature in all

C4HC composite films suggests the miscibility among the components. The XRD results

illustrates that the crystalline nature of CS decreases for the C4HC films due to the existence of

4HC. The results of water contact angle study showed that contact angle decreased with addition

of 4HC onto the CS indicating the films were hydrophilic in nature and affinity towards water

increased in C4HC films when compared to pure CS. Further study can be extended to the

application level by performing the different application oriented instrumental characterizations. It

can be expected that, the best properties of C4HC composite films were recorded in the study may

play a vital role in food packaging and biomedical applications.

ACKNOWLEDGEMENT

The author would like to acknowledge sincere gratitude to University Science Instrument Centre,

Karnatak University, Dharwad, Karnataka, India, for providing Characterization facility to study

properties.

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